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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (2): 165-172.doi: 10.3736/jcim20110209

• Original Experimental Research • Previous Articles     Next Articles

Dose-dependent effects of daidzein in regulating bone formation through estrogen receptors and peroxisome proliferator-activated receptor γ

Lei Bao, Shi-en Zou, Shao-fen Zhang()   

  1. Department of Gynaecology, Obstetrics and Gynaecology Hospital of Fudan University, Shanghai 200011, China
  • Received:2010-08-12 Accepted:2010-09-17 Online:2011-02-20 Published:2011-02-15
  • Contact: Zhang Shao-fen E-mail:zhangshaofen@163.com

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Objective: To investigate different doses of daidzein (DAI) in regulating bone formation of osteoblasts, and the regulating mechanisms of estrogen receptors (ERs) and peroxisome proliferator-activated receptor γ (PPARγ) in bone formation.
Methods: Human fetal osteoblasts (hFOBs) incubated without any treatment were served as controls (control group). The hFOBs were exposed to DAI of 10-9, 10-7 and 10-5 mol/L for 72 h, and to β-estradiol-17-valerate (E2) of 10 -8 mol/L as positive control, respectively. Methyl thiazolyl tetrazolium assay was employed to determine the proliferation status of osteoblasts, and 4-nitrophenyl phosphate disodium salt (PNPP) method was employed to determine the activity of alkaline phosphatase (ALP). ER antagonist ICI 182780 (ICI), ERα-selective antagonist methyl-piperidino-pyrazole (MPP) and irreversible PPARγ antagonist GW9662 (GW) were used to block the corresponding receptor, while hFOBs were exposed to E2 or different concentrations of DAI for 48 h. MTT assay and PNPP method were used respectively to determine the proliferation status and ALP activity of osteoblasts cultured in vitro.
Results: The osteoblast proliferation rate decreased progressively as the dose of DAI increased. Compared with the controls, the osteblast proliferation rate in the DAI 10-9 mol/L group increased significantly, while DAI 10 -5 mol/L group decreased significantly (P<0.05). ALP level decreased progressively as the dose of DAI increased, but there was no significant difference between groups (P>0.05). When ERs were blocked by ICI, proliferation rates in the E2 group and DAI 10-9, 10-7 and 10-5 mol/L groups were 88.16%, 76.30%, 81.18% and 83.19% respectively, which were all significantly lower than before (P < 0.05). After ERα was blocked by MPP alone, proliferation rates in E2 group and DAI 10-9, 10-7 and 10-5 mol/L groups were 69.78%, 63.31%, 70.71% and 78.43%, respectively, which were also significantly lower than before (P<0.05). ALP level in the DAI 10-9 mol/L group decreased significantly when ERα was blocked alone. When PPARγ inhibitor GW was added to the culture system, proliferation rates in E2 group and DAI 10-9, 10-7 and 10-5 mol/L groups were 103.14%, 96.99%, 112.88% and 122.22%, respectively. Compared with before, proliferation rates in DAI 10-7 and 10-5mol/L groups increased significantly (P<0.05), and ALP level increased significantly (P<0.05) in the DAI 10-5mol/L group.
Conclusion: DAI shows a biphasic effect on osteoporosis, whereby the effect is dose-dependent; a low-dose DAI stimulates proliferation of osteoblasts, while a high-dose DAI inhibits proliferation of osteoblasts. Low-dose DAI mainly acts on ERs, whereas high-dose DAI mainly acts on PPARγ to inhibit proliferation of osteoblasts and to some extent, acts on ERs to promote the proliferation of osteoblasts.

Key words: osteoporosis, postmenopausal, daidzein, receptors, estrogen, peroxisome proliferator-activated receptors

Figure 1

Dose-dependent effects of DAI on hFOBs A: The proliferation status of hFOBs was determined by MTT assay; data are expressed by the value of optical density (OD) at 570 nm. B: The ALP activity was determined by PNPP method; data are expressed by the value of OD at 405 nm. All the data are expressed as x±s, n=8. *P<0.05, vs control group; △P<0.05, vs E2 group; ▲P<0.05, vs D9 group. DAI: daidzein; hFOBs: human fetal osteoblasts; MTT: methyl thiazolyl tetrazolium; ALP: alkaline phosphatase; PNPP: 4-nitrophenyl phosphate disodium salt; E2: 10-8 mol/L β-estradiol-17-valerate; D9: 10-9 mol/L DAI; D7: 10-7 mol/L DAI; D5: 10-5 mol/L DAI."

Figure 2

Effects of blocked ERs on hFOBs A: The proliferation status of hFOBs was determined by MTT assay; data are expressed by the percentage of cell proliferation rate. B: The ALP activity was determined by PNPP method; data are expressed by the value of optical density (OD) at 405 nm. All the data are expressed as x±s, n=8. □P<0.05, vs before blocking group; ■P<0.05, vs +ICI group; ☆P<0.05, vs DAI 10-9 mol/L+ICI group. DAI: daidzein; ERs: estrogen receptors; hFOBs: human fetal osteoblasts; MTT: methyl thiazolyl tetrazolium; ALP: alkaline phosphatase; PNPP: 4-nitrophenyl phosphate disodium salt; E2: 10-8 mol/L β-estradiol-17-valerate; D9: 10-9 mol/L DAI; D7: 10-7 mol/L DAI; D5: 10-5 mol/L DAI; ICI: antiestrogen ICI 182780; MPP: ERα-selective antagonist, methyl-piperidino-pyrazole."

Figure 3

Effects of blocked PPARγ on hFOBs A: The proliferation status of hFOBs determined by MTT assay; data are expressed by the value of optical density (OD) at 570 nm. B: The ALP activity determined by PNPP method; data are expressed by the value of OD at 405 nm. All the data are expressed as x±s, n=8. □P<0.05, vs before blocking group. DAI: daidzein; PPARγ: peroxisome proliferator-activated receptor γ; hFOBs: human fetal osteoblasts; MTT: methyl thiazolyl tetrazolium; ALP: alkaline phosphatase; PNPP: 4-nitrophenyl phosphate disodium salt; E2: 10-8 mol/L β-estradiol-17-valerate; D9: 10-9 mol/L DAI; D7: 10-7 mol/L DAI; D5: 10-5 mol/L DAI; ICI: antiestrogen ICI 182780; MPP: ERα-selective antagonist, methyl-piperidino-pyrazole; GW: PPARγ antagonist GW9662."

Figure 4

Effects of ERs and PPARγ on proliferation rate of hFOBs Data are expressed by the percentage of cell proliferation rate. DAI: daidzein; D9: 10-9 mol/L DAI; D7: 10-7 mol/L DAI; D5: 10-5 mol/L DAI; ERs: estrogen receptors; PPARγ: peroxisome proliferator-activated receptor γ; hFOBs: human fetal osteoblasts."

[1] Piao JH, Pang LP, Liu ZH, Xiang Q, Su N, Pan ZA, Guo YC, Li FF, Li FG, Liu JP, Chen ZJ, Wang XM, Hu YW, Zhang YQ, Wang XH, Cheng XG . Chinese population, the diagnostic criteria of primary osteoporosis and the incidence of osteoporosis in China[J]. Zhonguo Gu Zhi Shu Song Za Zhi, 2002,8(1):1-7
朴俊红, 庞莲萍, 刘忠厚, 向青, 苏南, 潘子昂, 郭亦超, 李芳芳, 李扶刚, 刘京萍, 陈仲景, 王晓敏, 扈英伟, 张燕晴, 王晓红, 程晓光 . 中国人口状况及原发性骨质疏松症诊断标准和发生率[J]. 中国骨质疏松杂志, 2002,8(1):1-7
[2] Zou SE, Zhang SF, Jiang L, Zhang R, Zhang J . The dose-dependent effect of daidzein on osteoporosis in the ovariectomized rats[J]. Zhonguo Gu Zhi Shu Song Za Zhi, 2008,14(3):177-182
doi: 10.3969/j.issn.1006-7108.2008.03.008
邹世恩, 张绍芬, 蒋莉, 张锐, 张婧 . 大豆苷原防治去卵巢大鼠骨质疏松症的量效作用[J]. 中国骨质疏松杂志, 2008,14(3):177-182
doi: 10.3969/j.issn.1006-7108.2008.03.008
[3] Nuttall ME, Gimble JM . Controlling the balance between osteoblastogenesis and adipogenesis and the consequent therapeutic implications[J]. Curr Opin Pharmacol, 2004,4(3):290-294
doi: 10.1016/j.coph.2004.03.002
[4] Ricketts ML, Moore DD, Banz WJ, Mezei O, Shay NF . Molecular mechanisms of action of the soy isoflavones include activation of promiscuous nuclear receptors. A review[J]. J Nutr Biochem, 2005,16(6):321-330
doi: 10.1016/j.jnutbio.2004.11.008
[5] Bitto A, Burnett BP, Polito F, Marini H, Levy RM, Armbruster MA, Minutoli L, Di Stefano V, Irrera N, Antoci S, Granese R, Squadrito F, Altavilla D . Effects of genistein aglycone in osteoporotic, ovariectomized rats: a comparison with alendronate, raloxifene and oestradiol[J]. Br J Pharmacol, 2008,155(6):896-905
doi: 10.1038/bjp.2008.305
[6] Sharan K, Siddiqui JA, Swarnkar G, Maurya R, Chattopadhyay N . Role of phytochemicals in the prevention of menopausal bone loss: evidence from in vitro and in vivo, human interventional and pharmacokinetic studies[J]. Curr Med Chem, 2009,16(9):1138-1157
doi: 10.2174/092986709787581806
[7] Beresford JN, Bennett JH, Devlin C, Leboy PS, Owen ME . Evidence for an inverse relationship between the differentiation of adipocytic and osteogenic cells in rat marrow stromal cell cultures[J]. J Cell Sci, 1992,102(Pt 2):341-351
[8] Pei L, Tontonoz P . Fat’s loss is bone’s gain[J]. J Clin Invest, 2004,113(6):805-806
doi: 10.1172/JCI21311
[9] Dang ZC, Lowik C . Dose-dependent effects of phytoestrogens on bone[J]. Trends Endocrinol Metab, 2005,16(5):207-213
doi: 10.1016/j.tem.2005.05.001 pmid: 15922618
[10] Nakai M, Cook L, Pyter LM, Black M, Sibona J, Turner RT, Jeffery EH, Bahr JM . Dietary soy protein and isoflavones have no significant effect on bone and a potentially negative effect on the uterus of sexually mature intact Sprague-Dawley female rats[J]. Menopause, 2005,12(3):291-298
doi: 10.1097/01.GME.0000146109.50235.DO
[11] Kreijkamp-Kaspers S, Kok L, Grobbee DE, de Haan EH, Aleman A, Lampe JW, van der Schouw YT . Effect of soy protein containing isoflavones on cognitive function, bone mineral density, and plasma lipids in postmenopausal women: a randomized controlled trial[J]. JAMA, 2004,292(1):65-74
[12] Gallagher JC, Satpathy R, Rafferty K, Haynatzka V . The effect of soy protein isolate on bone metabolism[J]. Menopause, 2004,11(3):290-298
doi: 10.1097/01.GME.0000097845.95550.71 pmid: 15167308
[13] Arts J, Kuiper GG, Janssen JM, Gustafsson JA , Löwik CW, Pols HA, van Leeuwen JP.Differential expression of estrogen receptors α and β mRNA during differentiation of human osteoblast SV-HFO cells[J]. Endocrinology, 1997,138(11):5067-5070
doi: 10.1210/endo.138.11.5652
[14] Lindberg MK, Movérare S, Skrtic S, Gao H, Dahlman-Wright K, Gustafsson JA, Ohlsson C . Estrogen receptor(ER)-β reduces ERα-regulated gene transcription, supporting a “ying yang” relationship between ERα and ERβ in mice[J]. Mol Endocrinol, 2003,17(2):203-208
doi: 10.1210/me.2002-0206
[15] Maruyama S, Fujimoto N, Asano K, Ito A . Suppression by estrogen receptor β of AP-1 mediated transactivation through estrogen receptor α[J]. J Steroid Biochem Mol Biol, 2001,78(2):177-184
doi: 10.1016/S0960-0760(01)00083-8
[16] Keller H, Givel F, Perroud M, Wahli W . Signaling cross-talk between peroxisome proliferator-activated receptor/retinoid X receptor and estrogen receptor through estrogen response elements[J]. Mol Endocrinol, 1995,9(7):794-804
[17] Wang X, Kilgore MW . Signal cross-talk between estrogen receptor alpha and beta and the peroxisome proliferator-activated receptor gamma1 in MDA-MB-231 and MCF-7 breast cancer cells[J]. Mol Cell Endocrinol, 2002,194(1-2):123-133
doi: 10.1016/S0303-7207(02)00154-5
[18] Dang ZC , Löwik CW .The balance between concurrent activation of ERs and PPARs determines daidzein-induced osteogenesis and adipogenesis[J]. J Bone Miner Res, 2004,19(5):853-861
doi: 10.1359/jbmr.040120
[19] Dang ZC, Audinot V, Papapoulos SE, Boutin JA , Löwik CW .Peroxisome proliferator-activated receptor γ(PPARγ) as a molecular target for the soy phytoestrogen genistein[J]. J Biol Chem, 2003,278(2):962-967
doi: 10.1074/jbc.M209483200
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